Molecular isolation and identification of Mycobacterium avium subsp.hominissuis in Didelphis virginiana from Hidalgo,Mexico

Objective:To isolate and identify the exact species of the genus Mycobacterium from Didelphis(D.)virginiana,and the direct implications of this bacterium to public health and veterinary medicine.Methods:Thirty-one D.virginiana were captured and necropsied in Hidalgo,Mexico.Tissue samples were collected to culture mycobacteria present and examine individual specimens’histopathology.Mycobacterium identification was obtained through the application of amplification and sequencing of 16S rDNA techniques.Results:Three strains were isolated and identified as Mycobacterium(M.)avium subsp.hominissuis by utilizing M.avium complexspecific primers.Granulomatous lesions were observed in the subpleural zone(granuloma gradeⅡ)and bronchial(granuloma gradeⅠ)of the lungs of D.virginiana with positive isolation.Conclusions:Three strains of M.avium subsp.hominissuis,from lung tissue samples of D.virginiana were identified.This subspecies of M.avium has important implications in public health and veterinary medicine.

河南省南召县种植车厘子(Prunus avium L.)气候因子适宜性分析

利用年均温度、降雨量、日照时数、日照百分率和湿热指数等气候因子对河南省南召县种植车厘子的适宜性进行了分析研究.结果表明:南召县2011—2018年期间年平均气温(15.4℃)明显高于车厘子适宜的年均气温(10~12℃);年平均降雨量(743.7 mm)在车厘子生长发育对水分要求的适宜范围之内(600~800 mm);年均日照时数(1693.5 h)明显低于适宜的日照时数(2600~2800 h);1986—2002年的湿热指数与国内外车厘子主产区相比明显偏高.从大的气候范围来看,与国内外车厘子主产区相比,在南召县种植车厘子可能有树木生长旺盛、花芽形成不足、果实品质欠佳的风险.但充分利用东部浅山丘陵区等局部地区的小气候发展车厘子的种植是可行的.建议结合当地的温度、降雨量、日照时数和日照百分率等综合气候因子,选择合适的品种类型进行种植.

High prevalence of viable Mycobacterium avium subspecies paratuberculosis in Crohn's disease

AIM:To examine the detection rate of viable Mycobacterium avium subspecies paratuberculosis(MAP) in patients with inflammatory bowel disease [Crohn's disease(CD) and ulcerative colitis(UC)].METHODS:Thirty patients with CD(15 with at least one NOD2/CARD15 mutation),29 with UC,and 10 with no inflammatory bowel disease(IBD).were tested for MAP by polymerase chain reaction(specific IS900 fragment) and blood culture.RESULTS:MAP DNA was detected in all original blood samples and 8-wk blood cultures(CD,UC and non-IBD).Positive MAP DNA status was confirmed by dot blot assays.All 69 cultures were negative by acid-fast Ziehl-Neelsen staining.Viable MAP,in spheroplast form,was isolated from the 18-mo blood cultures of all 30 CD patients,one UC patient,and none of the non-IBD controls.No association was found between positive MAP cultures and use of immunosuppressive drugs or CDassociated single nucleotide polymorphisms.CONCLUSION:MAP is widely present in our area and MAP DNA can be recovered from the blood of CD,UC and non-IBD patients.However,MAP spheroplasts were only found in CD patients.

The relative frequency of Mycobacterium tuberculosis and Mycobacterium avium infections in HIV positive patients,Ahvaz,Iran

Objective:To estimate the prevalence of Mycobacterium tuberculosis(M.tuberculosis) and Mycobacterium avium(M.avium) infections in HIV -positive patients suspected to have puimonar) and extrapulmonary mycobacterial co-infection using PCR technique.Methods: Totally 50 samples comprising sputum,pleural fluid and CSF taken from HIV positive patients suspected to have mycobacterial infection,were processed.The demographic information and results of acid fast staining and culture were recorded for each patient.The PCR for detecting of M.tuberculosis comprised of specific primers targeting IS6110 gene sequence.For detecting of M.avium,PCR with primers that amplifies the mig gene were used.Results:From 50 samples processed,45 were sputum(90%),3 pleural fluid(6%) and 2 CSK(4%).In total.8(16%) were culture positive,7 had positive acid fast staining 114%) and 13 samples(26%)were positive using PCR technique.All the positive samples were sputum and belonged to patients with pulmonary infection.01 these.9 were positive for M.tuberculosis(69.2%) and 4 were identified as 1/.avium (30.8%).which 2 out of 13 positive samples showed mixed infections by both mycobacteria. Conclusions:The PCR shows the highest detection rate(26%) of mycobacteria compared with culture and acid last staining.The majority of infections were with M.tuberculosis(18%) and this shows the importance of this mycobacterial co-infection in HIV positive patients in the region of study.

On deaf ears, Mycobacterium avium paratuberculosis in pathogenesis Crohn's and other diseases

The historic suggestion that Mycobacterium avium subsp. paratuberculosis(Map) might be a zoonotic pathogen was based on the apparent similarity of lesions in the intestine of patients with Crohn's disease(CD) with those present in cattle infected with Map, the etiological agent of Johne's disease. Reluctance to fully explore this possibility has been attributed to the difficulty in demonstrating the presence of Map in tissues from patients with CD. Advances in technology have resolved this problem and revealed the presence of Map in a significant proportion of patients with CD and other diseases. The seminal finding from recent investigations, however, is the detection of Map in healthy individuals with no clinical signs of disease. The latter observation indicates all humans are susceptible to infection with Map and lends support to the thesis that Map is zoonotic, with a latent stage of infection similar to tuberculosis, where infection leads to the development of an immune response that controls but does not eliminate the pathogen. This clarifies one of the reasons why it has been so difficult to document that Map is zoonotic and associated with the pathogenesis of CD and other diseases. As discussed in the present review, a better understanding of the immune response to Map is needed to determine how infection is usually kept under immune control during the latent stage of infection and elucidate the triggering events that lead to disease progression in the natural host and pathogenesis of CD and immune related diseases in humans.

Mycobacterium avium subspecies paratuberculosis in the causation of Crohn's disease

Mycobacterium avium subspecies paratuberculosis(MAP),originally called Johne’s bacillus was firstdescribed from Germany in 1895 as the cause of achronic inflammatory disease of the intestine in acow.As the 20th century progressed,clinical andsub-clinical MAP infection in farm animals inWestern Europe appeared to become moreprevalent.Among the early reviews available

Mycobacterium avium subspecies paratuberculosis in the etiology of Crohn's disease, cause or epiphenomenon?

The origin of inflammatory bowel disease is unknown. Attempts have been made to isolate a microorganism that could explain the onset of inflammation, but no pathological agent has ever been identified. Johne's disease is a granulomatous chronic enteritis of cattle and sheep caused by Mycobacterium avium subspecies paratuberculosis(MAP) and shows some analogies with Crohn's disease(CD). Several studies have tried to clarify if MAP has a role in the etiology of CD. The present article provides an overview of the evidence in favor and against the "MAP-hypothesis", analyzing the methods commonly adopted to detect MAP and the role of antimycobacterial therapy in patients with inflammatory bowel disease. Studies were identified through the electronic database, MEDLINE, and were selected based on their relevance to the objective of the review. The presence of MAP was investigated using multiple diagnostic methods for MAP detection and in different tissue samples from patients affected by CD or ulcerative colitis and in healthy controls. On the basis of their studies, several authors support a close relationship between MAP and CD. Although increasing evidence of MAP detection in CD patients is unquestionable, a clear etiological link still needs to be proven.

Pulmonary <i>Mycobacterium avium </i>Complex Disease Requiring Differentiation from Recurrence of Lung Cancer during the Follow-Up Period for Lung Cancer

Case 1 was a 49-year-old woman who visited with a dry cough. She had an underlying disease of lung adenocarcinoma and received cancer immunotherapy because of an ALK-positive response and several cancer chemotherapies. The clinical effect was a complete response. Chest CT was performed because of continuous dry cough, and a new tumor shadow was recognized in the lingula portion of the left upper lobe. We performed CT-guided lung biopsy and could aspirate pus-fluid. The culture test for acid-fast bacilli was positive and the causative microorganism was identified as Mycobacterium avium by the DDH method. The final diagnosis was pulmonary abscess due to M. avium. Treatment using combined chemotherapy including CAM was performed and a good clinical response was obtained. Case 2 was a 67-year-old man who had a past history of surgical resection of lung adenocarcinoma eight and two years ago and received several cancer chemotherapies and radiation therapy. Because a new nodular shadow appeared in the right middle lobe one year ago and showed strong positivity on PET/CT, surgical resection was performed with the suspected recurrence of lung cancer. Subsequently, the histological diagnosis was epithelioid granuloma and a culture test of acid-fast bacilli was positive, with the identification of Mycobacterium intracellulare by the DDH method. Combined chemotherapy was not performed because the lesion was completely resected. Afterwards, a new nodular shadow appeared in the left lower lobe again and bronchoscopy was performed. Because M. intracellulare was isolated from the local specimen, we diagnosed the patient with recurrence of pulmonary MAC disease and combined chemotherapy including CAM was performed for one year. Finally, the nodular lesion disappeared. It is difficult to differentiate pulmonary MAC disease from lung cancer. Therefore, careful follow-up of patients with lung cancer while keeping in mind the possible complication of pulmonary MAC disease is necessary.

Concurrent pulmonary Mycobacterium avium complex (MAC) infection and active Hürthle cell thyroid carcinoma: is there a connection?

We present two cases of pulmonary MAC infection in women with Hürthle cell thyroid carcinoma. Both cases were asymptomatic octogenarian women with active Hürthle cell thyroid carcinoma and prolonged periods of hypothyroidism prior to diagnosis of pulmonary MAC. Mycobacterium avium complex has never been reported in association with any type of thyroid cancer, specifically Hürthle cell carcinoma. A review of the literature and possible associations between the two are discussed in this article.

Mycobacterium avium Paratuberculosis (MAP) and Cytomegalovirus (CMV) Are Frequently Detected in the Saliva of Patients Recently Diagnosed with Crohn Disease (CD) Whereas Oral Propionibacterium Acnes (PA) or Methylacetate (MA) in Their Breath Is Rare

We investigated the presence of MAP, CMV, or Epstein-Barr virus (EBV) in the saliva of patients presenting with periodontal disease, gingivitis or oral aphthosis. We also measured methylacetate in their breath and looked for the presence of PA on their tongue. All patients were prospectively enrolled into a two-year cohort study. We compared the group presenting with CD versus the group without CD. 167 patients were enrolled. 24 patients presented with CD. CMV and MAP were found concomitantly in 67% of CD patients (17 patients) whereas only 1 patient (<0.1%;p < 0.001) has these infectious agents in the comparative group. PA was rare in CD (25% versus 70.6%;p < 0.001). MA was low, especially with an empty stomach (0.59 ppm +/? 1.45 versus 5.14 ppm +/? 8.97;p < 0.001). It was concluded that the association MAP + CMV was frequently found in oral lesions of CD at an early stage. Detection of PA, MAP and CMV, as well as breath test could be easily performed and may participate to the early diagnosis of CD.

Effects of Different Hormone Proportions on Differentiation and Regeneration of Prtmus avium L. Adventitious Buds

In order to screen the appropriate culture condition for the differentiation and regeneration of Prtmus avium L. adventitious buds, in this study, the effect of different hormone proportions on differentiation and regeneration of shoot tip explants were investigated using Gisela No. 5 and Gisela No. 6 as experimental materi- als. The results showed that, different hormone proportions had extremely significant effects （ P 〈0.01 ） on the differentiation rate of P. avium adventitious buds; the appropriate hormone proportions for Gisela No. 5 and Gisela No. 6 to induce dedifferentiation of adventitious buds were 6-BA 3.0 mg/L ＋ IBA 0.5 mg/L ＋ KT O. 1 mg/L and 6-BA 1.0 mg/L ＋ IBA 0.5 mg/L ＋ KT 0.2 rag/L, respectively. In addition, different hormone proportions had extremely significant effects （P 〈 0.01 ） on the regeneration coefficient and regeneration rate of P. avium adventitious buds; with the hormone proportion of 6-BA 1.0 mg/L ＋ IBA 1.0 mg/L ＋ KT 0.3mg/L, the number of regenerated adventitious buds reached the maximum for both varieties.

The 19 kDa Protein from <i>Mycobacterium avium subspecies paratuberculosis</i>Is a Glycolipoprotein

This study characterizes the 19 kDa protein expressed by Mycobacterium avium subspecies paratuberculosis (MAP) as a glycolipoprotein, providing the foundation for future experiments regarding its antigenicity and role in disease pathogenicity. We have previously shown that a 4.8 kb insert from MAP will produce a 16 kDa recombinant protein when expressed in Escherichia coli and 19 kDa recombinant protein when expressed in M. smegmatis (smeg19K). The difference of 3 kDa in size of these expressed proteins may be related to post translational modifications that occur in Mycobacterium species. We hypothesized that smeg19K is a glycolipoprotein since BLAST analysis revealed approximately 76% amino acid identity between the MAP 19 kDa protein and a known lipoglycoprotein, the 19 kDa protein of M. tuberculosis. This prediction was confirmed by the following positive staining of smeg19K with Sudan Black 4B, a postelectrophoresis dye used to stain for lipids. Smeg19K has also stained positively for glycosylation with the lectin concavalin A, a highly specific stain for mannose residues. As expected, treatment with tunicamycin (an antibiotic known to inhibit N-glycosylation) and treatment with deglycosylation assay (non-specific for mannose), showed no reduction in size of 19 kDa glycolipoprotein.

Specific Antigens to Distinguish <i>M. tuberculosis</i>from <i>M. avium</i>

To distinguish Mycobacterium tuberculosis from Mycobacterium avium, specific M. tuberculosis antigens had been studied for improving the early differential diagnosis effect of tuberculosis caused by different Mycobacterium. The rabbit anti-M. avium sera and anti-M. tuberculosis sera were analyzed for antibody-based reactivity by matrix-assisted laser desorption-ionization mass spectrometry (MALDI-TOF Mass) against M. tuberculosis proteins. The immunoreactive spots, which were attributed to the proteins HspX, GroES and CFP-10, were mostly located at 10 - 60 kDa and PI 4 - 6, subsequently Western blotting result proved that HspX and CFP-10 were specific to M. tuberculosis and ELISA testing result of 30 M. avium positive sera showed that GroES were cross-reactive to M. avium. Lastly, positive and negative tuberculosis reference sera and based on the mechanism of indirect ELISA, the specificity and the sensitivity of the methods targeting the antibodies HspX, GroES or CFP-10 were evaluated at 37% and 26%, 12% and 97%, 81% and 98%, respectively. The combination of these three antibody detection methods allowed to reached a specificity of 42%, and of 39% without taken into account of the method targeting the GroES antibody. Using proteomics approach, we found three M. tuberculosis specific antigens showed good potential in tuberculosis diagnosis, providing basic study for serodiagnosis of tuberculosis.

In Vitro Studies: Potensial Use of Dyospiros kaky, Avium grafeolens, Guazuma ulmifolia, Hibiscus sabdariffa as an Agent of Anti-cholesterol

God has made man and has prepared everything for human life.This universe was created for humans to be healthy.The lifestyle of humans in the millenial era has made circulatory disorders in the form of hypercholesterolemia.This study aims to determine the efficacy of herbal plants to reduce blood cholesterol levels.The method used is the in vitro method.Herbal plant used in this experiment Dyospiros kaky 50%,Avium grafeolens 20%,Guazuma ulmifolia 20%,Hibiscus sabdariffa 10%.This study showed that cholesterol level decreased from 200 mg/dL to 102 mg/100 g at dose 20 mg/dL,64.5 mg/100 g,at dose 40 mg/dL,and 94.2 mg/100 g,at dose 60 mg/dL.The best dose to decrease blood cholesterol was 40 mg/dL.This study indicated that Dyospiros kaki is an agent of anti-cholesterol.

Identification of Prophages within the Mycobacterium avium 104 Genome and the Link of Their Function Regarding to Environment Survival

Mycobacterium avium is an opportunistic bacterium associated with pathogenic behavior in both humans and animals. M. avium has evolved as a pathogen by having an environmental component in its life style. Prophages are the integrated viral forms in bacterium genome. They constitute about 10% - 20% of genome of many bacteria and they contribute to pathogenicity of microbes. We investigated whether the M. avium 104 genome contained prophages and evaluated the genes/proteins for putative functions. Three prophage genes were identified in the M. avium 104 database, and sequences were analyzed for specific motifs. The prophage sequences were then cloned in Mycobacterium smegmatis and the bacterial phenotype was evaluated in gain of function assays for environmental stresses, such as tolerance to extreme temperatures, UV light, biofilm formation and resistance to acid as well as macrophage survival. The results indicate that two of the prophage genes, MAV_0696 and MAV_2265, confer M. smegmatis with enhanced ability to produce biofilm. Using a Real-Time PCR, it was determined that MAV_0696 and MAV_2265 transcripts were upregulated upon biofilm formation by M. avium. The expression of MAV_2265 gene was significantly higher at all selected time points. In addition, the expression of MAV_2265 in M. smegmatis also led to significantly greater survival rate at pH 5.0 compared to the wild-type control. None of the other physical abilities were altered by overexpressing the prophage genes in M. smegmatis. In summary, we identified three prophage sequences in M. avium 104, from which two of them were found to be associated with biofilm formation and one with resistance to the acidic environment. Future studies will identify the mechanisms involved in the prophages function.

Mycobacterium avium subspecies paratuberculosis and its relationship with Crohn's disease

The hypothesis postulating that Mycobacterium avium paratuberculosis(MAP) is the cause of Crohn's disease(CD) has been circulating for many years.Advances in molecular techniques,such as polymerase chain reaction and culture methods,have enabled researchers to demonstrate that there is an association between MAP and CD.Recently,genome-wide association studies have identified novel susceptibility genes for CD,which are critical for generation of an adaptive immune response that is protective against intracellular pathogens,including M.tuberculosis infection.However,the role of MAP as a cause of CD suffered a setback with the report that administration of antimycobacterial therapy failed to lead to a sustained response in CD patients.Accordingly,this review sought neither to confirm nor refute this,but instead to survey recent literature on the role of MAP in CD.

Relationship between Crohn’s disease, infection with Mycobacterium avium subspecies paratuberculosis and SLC11A1 gene polymorphisms in Sardinian patients

AIM: To study the association between Crohn’s disease (CD), Mycobacterium avium subspecies paratubercu- losis (MAP), and genetic factors by examining the role of natural resistance-associated macrophage protein 1 (NRAMP1) gene polymorphisms (now SLC11A1) in Sar dinian patients with CD and controls. METHODS: Thirty-seven CD patients and 34 controls with no inflammatory bowel disease (IBD) were recruited at the University of Sassari after giving written con sent. Six SCL11A1 polymorphisms previously reported to be the most significantly associated with IBD were searched. M. paratuberculosis was identified by IS900 PCR and sequencing. Logistic regression was used to cal culate odds ratios (OR) for the associations among CD presence of MAP, and 6 loci described above. RESULTS: For the first time, a strong association was observed between polymorphisms at NRAMP1 locus 823C/T and CD. While CD was strongly associated withboth NRAMP1 and MAP, NRAMP1 polymorphisms and MAP themselves were not correlated. CONCLUSION: Combined with previous work on the NOD2/CARD15 gene, it is clear that the interplay of ge- netic, infectious, and immunologic factors in the etiology of CD is complex.

Mycobacterium avium subspecies paratuberculosis infects and multiplies in enteric glial cells

AIM: To establish the role of enteric glial cells during infection with Mycobacterium avium subspecies paratuberculosis (MAP) in Crohn's disease. METHODS: In order to establish the role of enteric glial cells during infection with M. avium subspecies paratuberculosis (MAP) in Crohn's disease, Map adhesion experiments on enteric glial cells were performed as well as expression analysis of Map sigma factors during infection. RESULTS: In this study, for the first time, we found a high affinity of MAP to enteric glial cells and we analyzed the expression of MAP sigma factors under different conditions of growth. CONCLUSION: The fact that Map showed a high affinity to the glial cells raises concerns about the complicated etiology of the Crohn's disease. Elucidation of the mechanisms whereby inflammation alters enteric neural control of gut functions may lead to novel treatments for Crohn's disease.

海藻提取液对甜樱桃(Prunus avium)生长和果实品质的影响

2017、2018年春季在山东烟台进行了田间试验,研究了不同浓度海藻(铜藻)提取物的叶面施用对甜樱桃(Prunus avium)植物生长和果实品质的影响。通过叶面喷施不同稀释浓度的海藻提取液和清水对照,对生长和果实营养成分的相关指标进行了检测。2017年所有处理中,海藻提取液稀释浓度为100倍时的效果最显著,该处理组的叶面积,横径,纵径,单果重,硬度,果肉厚度和甜樱桃果实品质均优于对照组,可溶性糖和抗坏血酸含量显著提高。2018年根据2017年的实验结果,设置了实验进行重复验证。虽因天气原因,供试果园减产,但处理组仍表现出提高樱桃生长和果实品质的趋势。

牛副结核病的病原学和防控研究进展

牛副结核病是一种由副结核分枝杆菌引起的主要影响反刍动物的慢性消耗性疾病,也是一种潜在的人兽共患病。该病可导致腹泻、渐进性消瘦,甚至死亡,增加生产成本,是影响全球奶牛业最严重的传染病之一。牛副结核病难以诊断,病原分离周期长,我国相关研究报道较少且存在误区。因为潜伏期长,患病动物在感染早期体液免疫水平低,排菌量低,导致细菌分离培养和抗体检测都很难做出诊断;即使最灵敏的实时荧光定量聚合酶链式反应(qPCR)检测方法也只能在部分感染动物的粪便样品中检出副结核分枝杆菌;皮肤变态反应试验和γ干扰素释放试验等方法易受其他结核分枝杆菌的影响,也难以普及。本文旨在提炼性的阐述牛副结核病的病原学研究进展,总结其流行病学和防控策略等信息,以期为该病的预防和控制工作提供参考。